117 THE CHYMOTRYPSIN INHIBITOR CARBOBENZYLOXY-LEUCINE TYROSINE-CHLOROMETHYLKETONE (zLYCK) INTERFERES WITH THE NEUTROPHIL RESPIRATORY BURST MEDIATED BY A SIGNALING PATHWAY INDEPENDENT OF PtdinsP2 BREAKDOWN AND CYTOSOLIC FREE CALCIUM
117 THE CHYMOTRYPSIN INHIBITOR CARBOBENZYLOXY-LEUCINE TYROSINE-CHLOROMETHYLKETONE (zLYCK) INTERFERES WITH THE NEUTROPHIL RESPIRATORY BURST MEDIATED BY A SIGNALING PATHWAY INDEPENDENT OF PtdinsP2 BREAKDOWN AND CYTOSOLIC FREE CALCIUM
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117 糜蛋白酶抑制剂碳苄氧基亮氨酸酪氨酸氯甲基酮 (zLYCK) 干扰由独立于 PtdinsP2 分解和胞质游离钙的信号通路介导的中性粒细胞呼吸爆发
DOI：
10.1203/00006450-199112000-00147
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发表时间：
1991
影响因子：
3.6
通讯作者：
A. Verhoeven
中科院分区：
文献类型：
--
作者：
A. Gervaix;G. Kessels;S. Suter;D. Lew;A. Verhoeven
本文研究了胰凝乳蛋白酶抑制剂cbz-亮氨酸-酪氨酸-氯甲基酮（2LYCK）对人中性粒细胞呼吸爆发激活途径的影响。在10 μM时，zLYCK显示出对由化学引诱物甲酰-甲硫氨酰-亮氨酰苯丙氨酸（FMLP）刺激的超氧化物产生的平行抑制作用以及对人中性粒细胞的糜蛋白酶样活性的抑制作用。相比之下，由佛波醇酯乙酸酯（PMA）诱导的超氧化物产生受zLYCK的影响最小。分析了FMLP触发的已知转导途径。zLYCK既不影响FMLP诱导的胞浆游离钙瞬变、肌醇1，4，5三磷酸的形成，也不影响PMA诱导的蛋白激酶c的47 kD底物的磷酸化。zLYCK不影响中性粒细胞蛋白激酶c的活性。用来自刺激的中性粒细胞的活性膜或在细胞树系统中测试的NADPH氧化酶的活性不被zLYCK抑制。我们的结论是：1）zLYCK通过抑制中性粒细胞的胰凝乳蛋白酶样蛋白酶抑制超氧化物的产生2）zLYCK通过不依赖于PtdInsP 2断裂和胞浆游离钙的途径抑制FMLP诱导的NADPH氧化酶的激活3）zLYCK可能被证明是表征其在中性粒细胞激活中的靶蛋白酶的有用探针。
The effects of cbz-leucine-tyrosine-chloromethylketone(2LYCK), an inhibitor of chymotrypsin, were investigated in the activation pathways of human neutrophil respiratory burst. At 10 μM zLYCK showed a parallel inhibition of superoxide production stimulated with the chemoattractant formyl-methionyl-leucyl phenylalanine (FMLP) and chymotrypsin-like activity ot human neutrophils. By contrast superoxide production induced by phorbol mirystate acetate (PMA) was minimally affected by zLYCK. The known transduction pathways triggered by FMLP were analyzed. zLYCK did not affect either FMLP-induced cytosolic free calcium transient, inositol 1,4,5 triphosphate formation nor the PMA-induced phosphorylation of the 47 kD substrate of protein kinase c. zLYCK did not affect the activity of protein kinase c extracted from neutrophils. The activity ot the NADPH oxidase tested with active membranes from stimulated neutrophils or in a cell-tree-system was not inhibited by zLYCK. We conclude that 1) zLYCK inhibits superoxide production through the inhibition of a chymotrypsin-like protease of the neutrophil 2) zLYCK inhibits FMLP-induced activation of NADPH oxidase through a pathway independent of PtdInsP2 breakdwon and cytosolic free calcium 3) zLYCK may prove an useful probe for the characterization of its target protease in neutrophil activation.